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产琥珀酸拟杆菌利用巴氏甲烷八叠球菌 227 作为乙酸盐清除剂进行琥珀酸发酵的适应性研究。

Adaptation of Methanosarcina barkeri 227 as acetate scavenger for succinate fermentation by Actinobacillus succinogenes.

机构信息

Division of EcoScience and Interdisciplinary Program of EcoCreative, Graduate School, Ewha Womans University, Seoul, 03760, South Korea.

Department of Life Science, Ewha Womans University, Seoul, 03760, South Korea.

出版信息

Appl Microbiol Biotechnol. 2020 May;104(10):4483-4492. doi: 10.1007/s00253-020-10494-2. Epub 2020 Mar 17.

Abstract

Acetate is the main by-product from microbial succinate production. In this study, we performed acetate removal by Methanosarcina barkeri 227 for succinate fermentation by Actinobacillus succinogenes 130Z. The acetoclastic methanogen M. barkeri requires similar environmental factors to A. succinogenes, and the conditions required for co-cultivation were optimized in this study: gas used for anaerobicization, strain adaptation, medium composition, pH adjustment, and inoculation time points. M. barkeri 227 was adapted to acetate for 150 days, which accelerated the acetate consumption to 9-fold (from 190 to 1726 mmol gDW day). In the acetate-adapted strain, there was a noticeable increase in transcription of genes required for acetoclastic pathway-satP (acetate transporter), ackA (acetate kinase), cdhA (carbon monoxide dehydrogenase/acetyl-CoA synthase complex), and mtrH (methyl-HSTP:CoM methyltransferase), which was not induced before the adaptation process. The activities of two energy-consuming steps in the pathway-acetate uptake and acetate kinase-increased about 3-fold. This acetate-adapted M. barkeri could be successfully applied to succinate fermentation culture of A. succinogenes, but only after pH adjustment following completion of fermentation. This study suggests the utility of M. barkeri as an acetate scavenger during fermentation for further steps towards genetic and process engineering.

摘要

醋酸盐是微生物琥珀酸生产的主要副产物。在本研究中,我们利用 Methanosarcina barkeri 227 去除琥珀酸发酵过程中的醋酸盐,所用菌株为 Actinobacillus succinogenes 130Z。产乙酸甲烷菌 M. barkeri 与 A. succinogenes 需要类似的环境因素,本研究对共培养的条件进行了优化:厌氧化使用的气体、菌株适应、培养基成分、pH 值调整和接种时间点。M. barkeri 227 适应醋酸盐需要 150 天,这将醋酸盐的消耗速度提高了 9 倍(从 190 增加到 1726mmol gDW 天)。在适应醋酸盐的菌株中,参与乙酸分解代谢途径的基因 satP(乙酸转运蛋白)、ackA(乙酸激酶)、cdhA(一氧化碳脱氢酶/乙酰辅酶 A 合成酶复合物)和 mtrH(甲基-HSTP:CoM 甲基转移酶)的转录明显增加,而在适应之前这些基因并未被诱导。途径中两个耗能步骤的酶活性——乙酸摄取和乙酸激酶——增加了约 3 倍。这种适应醋酸盐的 M. barkeri 可以成功应用于 A. succinogenes 的琥珀酸发酵培养,但只能在发酵完成后进行 pH 值调整。本研究表明,M. barkeri 可作为发酵过程中的醋酸盐清除剂,用于进一步的遗传和工艺工程。

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